用抗合成多肽抗体分析呼吸道HAdV六邻体保守区抗原表位

目的：分析人类腺病毒（HAdV)六邻体保守区氨基酸序列，合成保守区多肽并制备抗多肽抗体，以确定型间共同的特异性表位。方法：用ClustalX,Antheprot等软件，对HAdV六邻体进行分析，确定保守区并分析其抗原性。合成抗原性多肽，免疫家兔制备抗肽多克隆抗体。用间接免疫荧光法和Western blot法，检测所制备的抗肽抗体与病毒抗原的反应性。结果：根据HAdV-2六邻体的保守区合成9种肽段，分成3组免疫家兔，共获得3组抗肽抗体。所有的抗肽抗体在1：1000稀释度时，与3个型别的HAdV感染细胞裂解物中相对分子质量（Mr)约为116000的六邻体亚单位具有特异性反应。抗肽抗体对HAdV感染细胞有特征性荧光染色。3组抗肽抗体中，有7个抗体与3个型别的HAdV感染细胞均具有阳性染色反应。结论：合成的保守区多肽能够诱导抗肽抗体产生，诱导产生的抗体与HAdV-3、－4、－7具有免疫反应性。通过分析预测抗原表位并制备抗肽抗体是可行的。     

抗钠钙交换体特异位点抗体对心肌钠钙交换电流的激活作用及其与钙通道、钠泵的交叉反应

目的：通过制备抗钠钙交换体（NCX）特异位点（α1重复序列124HNFTAGDLGPSTIVGSAAFNMF145）抗体探讨激动NCX的新途径，同时观察该抗体对于细胞膜上其它主要的离子通道和转运蛋白是否具有交叉反应。方法：将按照上述序列合成的肽段作为抗原， 对健康Wistar大鼠进行连续9周的主动免疫，用SA-ELISA法检测其血清中抗体的滴度，并对抗体滴度大于1∶〖KG-*2〗640的样本血清进行IgGs的提取和纯化。用膜片钳全细胞电流记录方法观测在10 nmol/L、20 nmol/L和40 nmol/L时该抗肽抗体对大鼠心室肌细胞钠钙交换电流（INa/Ca）、L-型钙电流（ICa-L）、钠泵电流（INa/k pump）、钠电流（INa）、瞬时外向钾电流（Itko）、内向整流钾电流（Ik1）以及豚鼠延迟整流钾电流（IK）的影响。结果：该抗肽抗体对于内向和外向的INa/Ca电流成分均产生剂量依赖性地激活作用，同时对IL-Ca和INa/k pump也有一定程度的激动，对检测的其余电流无显著影响。结论：该抗NCX特异位点抗体可以激动大鼠心肌钠钙交换电流，并且与L-型钙通道和钠泵具有交叉反应。     

病毒感染大鼠心室肌细胞钙通道基因表达及功能改变的研究

目的：研究大鼠心室肌细胞感染柯萨奇病毒B3（CVB3）后L型钙通道mRNA表达量及其电生理特性的变化。 方法: 用CVB3感染培养的SD大鼠心室肌细胞，采用半定量逆转录-聚合酶链式反应技术，检测病毒感染心室肌细胞L型钙通道各亚单位mRNA表达量的变化；用全细胞膜片钳技术，观察病毒感染前后心室肌细胞L型钙电流（ICa-L）的变化。结果: 病毒感染组心室肌细胞L型钙通道α1和β亚单位mRNA的表达量显著高于正常对照组（4.00±0.07 vs 2.21±0.41, P<0.01; 2.06±0.06 vs 1.22±0.30, P<0.05），而α2/δ亚单位mRNA表达量的改变不明显（4.12±0.19 vs 4.13±0.27, P>0.05）；感染组细胞ICa-L的平均电流密度明显大于正常组细胞［（-8.66±0.99) pA/pF vs (-6.97±1.75) pA/pF, P<0.01］，且前者的电流-电压曲线下移，峰电流密度增加25.74%（P<0.05）。结论: CVB3感染心室肌细胞后，使其L型钙通道α1和β亚单位mRNA的表达量增加，ICa-L增大，可能是病毒感染导致心肌出现异常电生理活动的细胞和分子机制之一。     

鸡肺表面活性蛋白A的表达及其多克隆抗体的制备

目的表达鸡肺表面活性蛋白A(cSP-A)重组蛋白,并制备鼠抗cSP-A多克隆抗体。方法人工合成cSP-A的凝集素糖识别区(CRD)基因(cSP-A-CRD),亚克隆到pGEX-6P-1载体中进行原核表达,对表达产物进行鉴定。切胶纯化法纯化重组蛋白,并免疫小鼠制备抗cSP-A多克隆抗体,ELISA检测抗体效价,Western blot法、免疫荧光组织化学染色和ELISA鉴定该抗体的特异性和适用范围。结果获得鼠抗cSP-A血清,效价达到105;Western blot法结果表明多抗血清具有良好的特异性和反应性,并可用免疫荧光组织化学技术检测鸡肺上皮细胞表达的cSP-A和用间接ELISA检测鸡肺灌洗液中存在的水溶性cSP-A蛋白。结论成功表达了cSP-A重组蛋白并制备了小鼠抗cSP-A多克隆抗体。     

抗人转录因子TFDP3和TFDP1多肽抗体的制备

目的：制备兔抗人转录因子TFDP3、TFDP1的多克隆抗体,并鉴定其反应性及特异性。方法：选择TFDP3和TFDP1差异较大肽段,利用人工合成多肽免疫家兔,采用真核表达载体瞬时转染HEK293细胞表达TFDP3和TFDP1真核蛋白,通过Western blot分析抗体的反应性和特异性。结果：通过氨基酸序列比对选择TFDP3 17-26,TFDP1 17-32氨基酸残基合成多肽,制备了兔抗人TFDP3、TFDP1多肽抗体,Western blot检测显示,抗TFDP3抗体可以特异性识别TFDP3,而抗TFDP1抗体既可识别TFDP1又可与TFDP3结合。结论：制备了兔抗人TFD31和TFDP1的多肽抗体,且TFDP3抗体与TFDP1无交叉反应。     

大鼠胚心室肌心房肽样物质定位的研究

本文用免疫组织化学方法研究了大鼠胚(13～19天)心房肽样免疫反应物质在心室肌细胞的定位,并在电镜下观察了大鼠胚心室肌细胞的超微结构。所得结果表明,心房肽类物质不仅存在于心房肌,亦在心室肌细胞内。含心房肽样免疫反应物质的细胞主要位于左心室,尤其是靠内膜侧的小梁状或网格状结构内。左右心室壁内这种细胞很少。心室肌细胞内所含心房肽样免疫反应颗粒的数量较同期心房肌细胞少。在电子显微镜下,见部分心室肌细胞含有与心房特殊颗粒(含心房肽类物质)相似的电子致密颗粒,它们大多靠近高尔基复合体。部分细胞不含这种颗粒,前者可能为心房肽样免疫反应阳性细胞,后者可能为阴性细胞。     

花生四烯酸对家兔心室肌细胞动作电位和L-型钙电流的影响

目的：研究花生四烯酸(AA)对家兔单个心室肌细胞动作电位和L-型钙电流的影响。方法：酶解法分离家兔单个心室肌细胞，用全细胞膜片钳技术记录其动作电位和L-型钙电流。结果：①AA明显缩短动作电位时程（APD），而对静息电位（RP）和动作电位幅值（APA）无明显影响。②AA使L- 型钙电流峰电流密度从(10.21±3.15) PA/PF减少到（6.53±2.17）PA/PF(n=6,P＜0.05）,I-V关系曲线上移，其形状和峰值电压保持不变。结论：花生四烯酸能抑制L-型钙电流，缩短动作电位时程，这可能是其心血管作用的重要机制之一。     

心肌肽素对大鼠心室肌细胞瞬时外向钾电流的影响

目的 研究新药心肌肽素对大鼠心室肌细胞瞬时外向钾电流(Ito)的影响及其在离子通道水平的作用机制。方法 用急性酶解法分离大鼠心室肌细胞及标准的全细胞膜片钳技术，观察不同浓度的心肌肽素对Ito的影响。结果 心肌肽素呈浓度依赖性抑制大鼠心室肌细胞Ito，心肌肽素浓度(mg/L)为10、50、100、250和500 时分别使Ito降低(%)4、13、22、32和38。心肌肽素50 mg/L使Ito电流密度-电压曲线下移，但不改变曲线的形状，也不改变Ito稳态激活曲线。结论 心肌肽素抑制大鼠心室肌细胞Ito，可能是其抗心律失常作用的机制之一。     

抗血管紧张素Ⅱ受体Ⅰ型抗体对成年大鼠心室肌细胞内游离钙离子浓度的影响

目的：研究抗血管紧张素Ⅱ受体Ⅰ型（AT1受体）抗体对成年大鼠心室肌细胞内游离钙离子浓度（[Ca^2＋]．）的影响。方法：用AT1受体胞外第2肽段合成肽免疫Wistar大鼠，以硫酸铵沉淀法粗提血清抗体后再以亲和层析法纯化抗体，并以ELISA和Bradford的方法对抗体进行定量。以Fluo-3AM作为细胞内游离Ca^2＋示踪剂，通过激光共聚焦显微镜检测分离的成年大鼠心室肌细胞[Ca^2＋]．的变化，以判断抗体活性，分AngⅡ组，抗AT1受体抗体组，抗AT1受体抗体＋氯沙坦组和对照组。结果：AngⅡ刺激后引起[Ca^2＋]．快速上升至峰值，随后快速下降（157．4±52．5）％；而在抗AT1受体抗体刺激下，呈双相反应，[Ca^2＋]．快速上升至峰值，随后进入平台期（164．0±34.0）％，与AngⅡ组相比无统计学意义（P〉0．05）。其效应部分被氯沙坦阻断（79．7±13．2）％，与前两组相比差异有统计学意义（P〈0．05）。结论：抗AT1受体抗体可以引起大鼠心室肌细胞内[Ca^2＋]，浓度增加，呈快速的峰值和持续的平台期的双相反应，主要依赖于细胞内储存钙离子的释放。     

抗CMG2单克隆抗体的制备及其初步应用

目的制备鼠抗人毛细血管形态发生基因2(CMG2)单克隆抗体,并将其初步应用于胃癌组织和细胞系中CMG2表达的检测。方法重组表达CMG2胞外区肽段,免疫Balb/c小鼠,常规制备杂交瘤细胞,通过间接ELISA方法筛选目标克隆,制备小鼠腹水并经蛋白A亲和纯化获得相应抗体。间接ELISA法鉴定抗体的亚类类型及亲和力;抗原竞争性抑制实验鉴定抗体的特异性。用候选抗体建立流式细胞术、免疫荧光、免疫印迹及免疫组织化学方法,初步应用于胃癌组织和细胞系中CMG2表达的检测。结果共获得3株针对CMG2胞外区的杂交瘤细胞株,其中以8F3单克隆抗体的亲和力和特异性最好,亚型为Ig G1。8F3单抗能用于胃癌细胞株CMG2表达的流式细胞术和免疫荧光检测以及胃癌组织的免疫组织化学检测。结论成功制备出高效价高特异性的鼠抗人CMG2单克隆抗体,为CMG2的基础和临床应用研究提供了有用工具。     

Renal dysplasia and asplenia in two sibs

A family is reported in which two sibs, one male and the other female, both died within 24 hours of birth with enlarged polycystic kidneys. Postmortem histology in the second child showed gross renal dysplasia. In both children the pancreas was enlarged, nodular and cystic but the liver appeared macroscopically normal. In the second child, histological examination confirmed pancreatic fibrosis with cystic dilation of ducts, but showed portal fibrosis with bile duct proliferation in the liver.
This combination of findings is very reminiscent of those in a girl and her brother reported by Ivemark et al. (1959). The children reported here also showed absence or hypoplasia of the spleen, cardiac anomalies and other features of the Ivemark syndrome (Ivemark 1955), a quite different, usually sporadic, congenital disorder. It is suggested that the children described here have a distinct lethal congenital disorder, probably inherited in an autosomal recessive manner.     

Schizophrenia in a North Swedish geographical isolate, 1900–1977. Epidemiology, genetics and biochemistry

Over 200 schizophrenic patients belonging to three major and interrelated pedigree complexes have been investigated over the past 30 years in a North Swedish geographically isolated population, presently numbering about 6,000. An intensive investigation of a number of biochemical correlates and genetic markers in a few selected families belonging to one of the major pedigrees has indicated new strategies for the current research program.
Schizophrenia, as defined operationally, is significantly associated with decreased activities of two enzymes (1) blood platelet monoamine oxidase, (2) plasma dopamine-β-hydroxylase, and (3) with the genetic marker Gc² (group specific antigen). Both enzymes are subject to genetic variation. A positive score for linkage between schizophrenia and low plasma DBH activity has been calculated, but, so far, available data are insufficient for discrimination between linkage and partial contribution of genetically controlled low plasma DBH to the pathogenesis of the disease. Alternatively, both mechanisms could be involved.
As a model for continued research, schizophrenia is explained as based on a double dominant-recessive genotype (Aabb), representing a vulnerability which in about 50 % of cases develops into clinical schizophrenia. It is suggested that the dominant mutation (A) operates on or affects MAO activity, and that the recessive genotype (bb) is instrumental in low variates of DBH activity and very likely such variates within the normal range of physiological variation. Moreover, it is suggested that the combined effects of MAO- and DBH-reduced efficiency on the metabolism of e.g. dopamine could be an essential pathogenic mechanism for the schizophrenic illness which is segregating in this population.     

The dominant diseases of modernized societies as omega-3 essential fatty acid deficiency syndrome: Substrate beriberi

About 1900, modern food selection and processing caused widespread $\text{epidemics}$ of the B vitamin deficiency diseases of beriberi and pellagra which, for genetic reasons, often expressed as different diseases ranging from bowel and heart disease to dermatoses and psychoses. But the B vitamins merely help convert essential fatty acids (EFA) into the prostaglandin (PG) tissue regulators and it now turns out that, through hydrogenation, milling and selection of w3-poor southern foods, we have also been systematically depleting, by as much as 90%, a newly discovered trace Nordic EFA (w3) of special importance to primates and sole precursor of the PG3(4) series, even as a concurrent fiber deficiency increases body demand for EFA. Since substrate EFA is processed by many B vitamin catalysts, an EFA deficiency will mimic a $\text{panh}$ypovitaminosis B, i.e., a mixture of $\text{substrate}$ beriberi and $\text{substrate}$ pellagra resembling vitamin beriberi and pellagra but exhibiting as even more diverse $\text{endemic}$ disease. This would consitute a second stage of the Modern Malnutrition and explain why some workers now hold the dominant diseases of modermized societies to be new, nutritionally based, pellagraform yet lipid-related and to range, once again, from heart disease to psychosis. It is an assumption that our dominant diseases are unrelated to each other or are merely revealed by our diagnostic acumen and therapeutic success; and that hydrogenating millions of tons of food oils annually, to destroy the rancidity producing w3-EFA, is safe for $\text{primates}$. Extensive beriberiform disease is reported here in 32 typical cases taken from medical practice which responds strikingly to linseed oil supplements (60% w3-EFA) in confirmation of identical results in Capuchins.     

What will studies of Fulani individuals naturally exposed to malaria teach us about protective immunity to malaria?

There are an estimated over 200 million yearly cases of malaria worldwide. Despite concerted international effort to combat the disease, it still causes approximately half a million deaths every year, the majority of which are young children with Plasmodium falciparum infection in sub-Saharan Africa. Successes are largely attributed to malaria prevention strategies, such as insecticide-treated mosquito nets and indoor spraying, as well as improved access to existing treatments. One important hurdle to new approaches for the treatment and prevention of malaria is our limited understanding of the biology of Plasmodium infection and its complex interaction with the immune system of its human host. Therefore, the elimination of malaria in Africa not only relies on existing tools to reduce malaria burden, but also requires fundamental research to develop innovative approaches. Here, we summarize our discoveries from investigations of ethnic groups of West Africa who have different susceptibility to malaria.     

'Very sore nights and days': the child's experience of illness in early modern England, c.1580-1720

Sick children were ubiquitous in early modern England, and yet they have received very little attention from historians. Taking the elusive perspective of the child, this article explores the physical, emotional, and spiritual experience of illness in England between approximately 1580 and 1720. What was it like being ill and suffering pain? How did the young respond emotionally to the anticipation of death? It is argued that children’s experiences were characterised by profound ambivalence: illness could be terrifying and distressing, but also a source of emotional and spiritual fulfilment and joy. This interpretation challenges the common assumption amongst medical historians that the experiences of early modern patients were utterly miserable. It also sheds light on children’s emotional feelings for their parents, a subject often overlooked in the historiography of childhood. The primary sources used in this article include diaries, autobiographies, letters, the biographies of pious children, printed possession cases, doctors’ casebooks, and theological treatises concerning the afterlife.     

Guidelines for the Validation and Use of Immunoassays for Determination of Introduced Proteins in Biotechnology Enhanced Crops and Derived Food Ingredients

Recent advancements in agricultural biotechnology have created a need for analytical techniques to determine introduced proteins in crops enhanced through modern biotechnology techniques. These proteins are expressed in plant tissues and may be present in food ingredients. Immunoassays are ideally suited for protein detection and may be used as both quantitative and threshold methods. Microplate ELISA and lateral flow devices are two of the most commonly used immunoassay formats for agricultural biotechnology applications. This paper provides general background information and a discussion of criteria for the validation and application of immunochemical methods to the analysis of proteins introduced into plants and food ingredients using biotechnology methods. It is the result of a collaborative effort of members of the Analytical Environmental Immunochemical Consortium. This collaborative effort represents the combined expertise of several organizations to reach consensus on establishing guidelines for the validation and use of immunoassays. Further, the paper offers developers and users a consistent approach to adopting the technology as well as aid in producing accurate and meaningful results.     

Ultracryotomy: development and application (with examples from the yeast cytology) (author's transl)

The preparation steps usually necessary for obtaining ultrathin frozen sections of biological material (chemical prefixation, enclosing, cryoprotective treatment, freezing, sectioning, and post-staining the sections for transmission electron microscopy) are submitted to a critical analysis. The application of cryo-ultramicrotomy, in particularly for cytochemical purposes, is reviewed. Fundamental considerations of chemical prefixation and poststaining are supported by examples from yeast cytology. Furthermore, the efficiency of the cryo-ultramicrotomy (electron optical resolution of ultrastructural details) is demonstrated on yeast cells and protoplasts.     

HLA in North Colombia Chimila Amerindians

HLA-A,-B,-C,-DRB1 and -DQB1 alleles have been studied in Chimila Amerindians from Sabana de San Angel (North Colombian Coast) by using high resolution molecular typing. A frequent extended haplotype was found:HLA-A*24:02-B*51:10-C*15:02-BRB1*04:07-DQB1*03:02 (28.7%) which has also been described in Amerinndian Mayos Mexican population (Mexico, California Gulf, Pacific Ocean). Other haplotypes had already been found in Amerindians from Mexico (Pacific and Atlantic Coast), Peru (highlands and Amazon Basin), Bolivia and North USA. A geographic pattern according to HLA allele or haplotype frequencies is lacking in Amerindians, as already known. Also, five new extended haplotypes were found in Chimila Amerindians. Their HLA-A*24:02 high frequencies characteristic is shared with aboriginal populations of Taiwan; also, HLA-C*01:02 high frequencies are found in New Zealand Maoris, New Caledonians and Kimberly Aborigines from Australia. Finally, this study may show a model of evolutionary factors acting and rising one HLA allele frequency (-A*24:02), but not in others that belong to the same or different HLA loci.     

The universal muscular chamber. A basic unit of the genitourinary, cardiovascular, gastro-intestinal, skeletal, cutaneous, respiratory and other systems, endocameral hypertension; and spasm, the key to their idiopathic diseases and arthropathies

Starting with the integument, we see many organs are contractile sacs or multiples thereof, which tubes or bags constitute the major part of the entire body. Recognition of this basic unit and its characteristics sheds new light, individually and collectively, on many disorders previously considered unrelated. Muscular tears and perforations develop in the walls of these chambers, being no way peculiar to those organs, wherein, hydrochloric acid occurs. So, it is not necessary to explain the absence of excessive acid from patients who exhibit holes in the gastric, uterine, aortic, duodenal, rectal, pulmonary, retina, and other walls. Muscle, not acid is the great common factor relating idiopathic disorders in the gastrointestinal tract to each other and to similar diseases in other systems. When the units are linked together, the lesions tend to appear as arthropathies, i.e. at the joints. Rephrasing common-place observations, frees us from conventional, conceptual cul-de-sacs. An observation is only as good as its interpretation, so all possibilities must be considered, otherwise, we will remain blinded by our misconceptions.